COI barcoding can distinguish bisexual and parthenogenetic populations of Haemaphysalis longicornis in Japan: Revisiting methods with SNP analysis as another possible method

Haemaphysalis longicornis, the Asian long-horned tick, is an important vector for various infectious diseases, such as severe fever with thrombocytopenia syndrome (SFTS) and Japanese spotted fever. In this species, a triploid parthenogenetic reproductive form occurs along with a diploid bisexual for...

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Main Authors: Mizue Inumaru, Kentaro Itokawa, Ryo Matsumura, Kyoko Sawabe, Mamoru Watanabe, Haruhiko Isawa, Shinji Kasai, Yukiko Higa
Format: Article
Language:English
Published: Elsevier 2025-08-01
Series:International Journal for Parasitology: Parasites and Wildlife
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Online Access:http://www.sciencedirect.com/science/article/pii/S2213224425000483
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author Mizue Inumaru
Kentaro Itokawa
Ryo Matsumura
Kyoko Sawabe
Mamoru Watanabe
Haruhiko Isawa
Shinji Kasai
Yukiko Higa
author_facet Mizue Inumaru
Kentaro Itokawa
Ryo Matsumura
Kyoko Sawabe
Mamoru Watanabe
Haruhiko Isawa
Shinji Kasai
Yukiko Higa
author_sort Mizue Inumaru
collection DOAJ
description Haemaphysalis longicornis, the Asian long-horned tick, is an important vector for various infectious diseases, such as severe fever with thrombocytopenia syndrome (SFTS) and Japanese spotted fever. In this species, a triploid parthenogenetic reproductive form occurs along with a diploid bisexual form. Several approaches have been used to distinguish these two groups, including the presence/absence of males in the population, karyotyping, flow cytometry, and most recently, mitochondrial phylogeny. Mitochondrial gene (COI) barcoding has also been casually used, although its validity has not been investigated. In the present study, the validity of COI barcoding, genotyping nuclear markers (SNPs), and morphometrics was evaluated for distinguishing the reproductive forms of H. longicornis in Japan. Ticks were collected using the flagging method at two locations in Hyogo, Japan. DNA was extracted from ticks after photography, which was used for morphometric measurements. The DNA was used for COI barcoding by direct sequencing and genotyping SNPs in the nuclear genome. The resulting COI haplotypes were clustered into two distinct haplogroups, which represented different ploidy levels, corresponding to the different reproductive groups. Genotypes of nuclear SNPs supported that the individuals from each mitochondrial haplogroup belonged to distinct reproductive populations with different ploidy levels. Meanwhile, although significant differences were observed in multiple morphometric characteristics between these reproductive groups, large overlaps were generally evident in the distribution, indicating that morphological identification is not sufficient to distinguish the reproductive groups. This study suggested for the first time that COI barcoding and SNP genotyping are both convenient and reliable methods to distinguish the two reproductive forms of H. longicornis in Japan.
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spelling doaj-art-7faae0b1e3bd451e9b2635b7cb158a702025-08-20T03:44:58ZengElsevierInternational Journal for Parasitology: Parasites and Wildlife2213-22442025-08-012710108310.1016/j.ijppaw.2025.101083COI barcoding can distinguish bisexual and parthenogenetic populations of Haemaphysalis longicornis in Japan: Revisiting methods with SNP analysis as another possible methodMizue Inumaru0Kentaro Itokawa1Ryo Matsumura2Kyoko Sawabe3Mamoru Watanabe4Haruhiko Isawa5Shinji Kasai6Yukiko Higa7Department of Medical Entomology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, Japan; Corresponding author.Department of Medical Entomology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, JapanDepartment of Medical Entomology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, Japan; Graduate School of Agriculture, Meiji University, Higashimita 1-1-1, Tama, Kawasaki, Kanagawa, 214-8571, JapanDepartment of Medical Entomology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, Japan; Graduate School of Agricultural and Life Sciences, University of Tokyo, Yayoi 1-1-1, Bunkyo, Tokyo, 113-8657, JapanDepartment of Medical Entomology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, JapanDepartment of Medical Entomology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, JapanDepartment of Medical Entomology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, JapanDepartment of Medical Entomology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, JapanHaemaphysalis longicornis, the Asian long-horned tick, is an important vector for various infectious diseases, such as severe fever with thrombocytopenia syndrome (SFTS) and Japanese spotted fever. In this species, a triploid parthenogenetic reproductive form occurs along with a diploid bisexual form. Several approaches have been used to distinguish these two groups, including the presence/absence of males in the population, karyotyping, flow cytometry, and most recently, mitochondrial phylogeny. Mitochondrial gene (COI) barcoding has also been casually used, although its validity has not been investigated. In the present study, the validity of COI barcoding, genotyping nuclear markers (SNPs), and morphometrics was evaluated for distinguishing the reproductive forms of H. longicornis in Japan. Ticks were collected using the flagging method at two locations in Hyogo, Japan. DNA was extracted from ticks after photography, which was used for morphometric measurements. The DNA was used for COI barcoding by direct sequencing and genotyping SNPs in the nuclear genome. The resulting COI haplotypes were clustered into two distinct haplogroups, which represented different ploidy levels, corresponding to the different reproductive groups. Genotypes of nuclear SNPs supported that the individuals from each mitochondrial haplogroup belonged to distinct reproductive populations with different ploidy levels. Meanwhile, although significant differences were observed in multiple morphometric characteristics between these reproductive groups, large overlaps were generally evident in the distribution, indicating that morphological identification is not sufficient to distinguish the reproductive groups. This study suggested for the first time that COI barcoding and SNP genotyping are both convenient and reliable methods to distinguish the two reproductive forms of H. longicornis in Japan.http://www.sciencedirect.com/science/article/pii/S2213224425000483COI barcodingHaemaphysalis longicornisMorphometric analysisParthenogenesisSNP genotyping
spellingShingle Mizue Inumaru
Kentaro Itokawa
Ryo Matsumura
Kyoko Sawabe
Mamoru Watanabe
Haruhiko Isawa
Shinji Kasai
Yukiko Higa
COI barcoding can distinguish bisexual and parthenogenetic populations of Haemaphysalis longicornis in Japan: Revisiting methods with SNP analysis as another possible method
International Journal for Parasitology: Parasites and Wildlife
COI barcoding
Haemaphysalis longicornis
Morphometric analysis
Parthenogenesis
SNP genotyping
title COI barcoding can distinguish bisexual and parthenogenetic populations of Haemaphysalis longicornis in Japan: Revisiting methods with SNP analysis as another possible method
title_full COI barcoding can distinguish bisexual and parthenogenetic populations of Haemaphysalis longicornis in Japan: Revisiting methods with SNP analysis as another possible method
title_fullStr COI barcoding can distinguish bisexual and parthenogenetic populations of Haemaphysalis longicornis in Japan: Revisiting methods with SNP analysis as another possible method
title_full_unstemmed COI barcoding can distinguish bisexual and parthenogenetic populations of Haemaphysalis longicornis in Japan: Revisiting methods with SNP analysis as another possible method
title_short COI barcoding can distinguish bisexual and parthenogenetic populations of Haemaphysalis longicornis in Japan: Revisiting methods with SNP analysis as another possible method
title_sort coi barcoding can distinguish bisexual and parthenogenetic populations of haemaphysalis longicornis in japan revisiting methods with snp analysis as another possible method
topic COI barcoding
Haemaphysalis longicornis
Morphometric analysis
Parthenogenesis
SNP genotyping
url http://www.sciencedirect.com/science/article/pii/S2213224425000483
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